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Recombinant Expression of Cyclotides Using Split Inteins

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Split Inteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1495))

Abstract

Cyclotides are fascinating microproteins (≈30 residues long) present in several families of plants that share a unique head-to-tail circular knotted topology of three disulfide bridges, with one disulfide penetrating through a macrocycle formed by the two other disulfides and inter-connecting peptide backbones, forming what is called a cystine knot topology. Naturally occurring cyclotides have shown to posses various pharmacologically relevant activities and have been reported to cross cell membranes. Altogether, these features make the cyclotide scaffold an excellent molecular framework for the design of novel peptide-based therapeutics, making them ideal substrates for molecular grafting of biological peptide epitopes. In this chapter we describe how to express a native folded cyclotide using intein-mediated protein trans-splicing in live Escherichia coli cells.

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Author information

Authors and Affiliations

  1. Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA, 90089-9121, USA

    Krishnappa Jagadish & Julio A. Camarero

  2. Department of Chemistry, University of Southern California, Los Angeles, CA, 90089-9121, USA

    Julio A. Camarero

Authors
  1. Krishnappa Jagadish
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  2. Julio A. Camarero
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Corresponding author

Correspondence to Julio A. Camarero .

Editor information

Editors and Affiliations

  1. Institute of Biochemistry Department of Chemistry and Pharmacy, University of Muenster, Münster, Germany

    Henning D. Mootz

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Jagadish, K., Camarero, J.A. (2017). Recombinant Expression of Cyclotides Using Split Inteins. In: Mootz, H. (eds) Split Inteins. Methods in Molecular Biology, vol 1495. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6451-2_4

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  • DOI: https://doi.org/10.1007/978-1-4939-6451-2_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6449-9

  • Online ISBN: 978-1-4939-6451-2

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